User Guide: DuoSpray Ion Source for TripleTOF Systems Operator Guide

User Guide: DuoSpray Ion Source for TripleTOF Systems Operator Guide

DuoSpray

Ion Source

for TripleTOF

®

Systems

Operator Guide

RUO-IDV-05-0783-A June 2014

This document is provided to customers who have purchased AB Sciex equipment to use in the operation of such AB Sciex equipment. This document is copyright protected and any reproduction of this document or any part of this document is strictly prohibited, except as AB Sciex may authorize in writing.

Software that may be described in this document is furnished under a license agreement. It is against the law to copy, modify, or distribute the software on any medium, except as specifically allowed in the license agreement. Furthermore, the license agreement may prohibit the software from being disassembled, reverse engineered, or decompiled for any purpose. Warranties are as stated therein.

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AB Sciex’s sole and exclusive representations, warranties, and obligations. AB Sciex makes no other warranty of any kind whatsoever, expressed or implied, including without limitation, warranties of merchantability or fitness for a particular purpose, whether arising from a statute or otherwise in law or from a course of dealing or usage of trade, all of which are expressly disclaimed, and assumes no responsibility or contingent liability, including indirect or consequential damages, for any use by the purchaser or for any adverse circumstances arising therefrom.

For research use only. Not for use in diagnostic procedures.

The trademarks mentioned herein are the property of AB Sciex Pte. Ltd. or their respective owners.

AB SCIEX ™ is being used under license.

© 2014 AB Sciex Pte. Ltd.

Printed in Canada.

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SINGAPORE 739256

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Contents

Chapter 1 Ion Source Overview.................................................................................................................5

Related Documentation.....................................................................................................................................................5

Technical Support..............................................................................................................................................................5

Ion Source Components.....................................................................................................................................................6

Probes................................................................................................................................................................................7

TurboIonSpray Probe....................................................................................................................................................8

APCI Probe...................................................................................................................................................................8

Gas and Electrical Connections..........................................................................................................................................9

Ion Source Sense Circuit...................................................................................................................................................10

Source Exhaust System....................................................................................................................................................10

Chapter 2 Ion Source Installation.............................................................................................................12

Prepare for Installation....................................................................................................................................................12

Install the Probes.............................................................................................................................................................13

Connect the Ion Source Tubing........................................................................................................................................14

Install the Ion Source on the Mass Spectrometer.............................................................................................................14

Plumb the Ion Source for Sample Introduction with the TurboIonSpray Probe.................................................................15

Plumb the Ion Source for Sample Introduction with the APCI Probe................................................................................16

Chapter 3 Ion Source Optimization.........................................................................................................17

Sample Introduction.........................................................................................................................................................17

Method......................................................................................................................................................................17

Flow Rate...................................................................................................................................................................17

Sample Inlet Requirements........................................................................................................................................18

TurboIonSpray Probe Optimization..................................................................................................................................18

Flow Rate and Temperature.......................................................................................................................................19

Set Up the System......................................................................................................................................................22

Run the Method.........................................................................................................................................................22

Set the Starting Conditions........................................................................................................................................19

Optimize the TurboIonSpray Probe Position...............................................................................................................20

Optimize Source and Gas Parameters and Voltage....................................................................................................21

Optimize the Turbo Heater Temperature....................................................................................................................21

APCI Probe Optimization.................................................................................................................................................22

Set Up the System......................................................................................................................................................22

Run the Method.........................................................................................................................................................22

Set the Starting Conditions........................................................................................................................................23

Optimize Gas 2 and Curtain Gas Flow ......................................................................................................................23

Adjust the Position of the Corona Discharge Needle.................................................................................................23

Optimize the APCI Probe Position..............................................................................................................................24

Optimize the IonSpray

Voltage Floating ...............................................................................................................25

Optimize the APCI Probe Temperature.......................................................................................................................25

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Contents

Chapter 4 Ion Source Maintenance..........................................................................................................26

Clean the Probes..............................................................................................................................................................27

Remove the Ion Source....................................................................................................................................................28

Remove the Probe............................................................................................................................................................28

Clean the Electrode Tube.................................................................................................................................................29

Assemble the Probe Components....................................................................................................................................31

Adjust the Electrode Tip Extension...................................................................................................................................31

Replace the Corona Discharge Needle Tip.......................................................................................................................32

Replace the Corona Discharge Needle.............................................................................................................................33

Replace the Sample Tubing..............................................................................................................................................35

Chapter 5 Troubleshooting Tips...............................................................................................................36

Appendix A Principles of Operation—Ion Source...................................................................................39

TurboIonSpray Mode........................................................................................................................................................39

APCI Mode.......................................................................................................................................................................40

APCI Ionization Region....................................................................................................................................................43

Appendix B Source Parameters and Voltages.........................................................................................46

TurboIonSpray Probe Parameters.....................................................................................................................................46

APCI Probe Parameters....................................................................................................................................................47

Parameter Descriptions....................................................................................................................................................48

Probe Position..................................................................................................................................................................50

Solvent Composition........................................................................................................................................................50

Appendix C Consumables and Spares.....................................................................................................52

Revision History........................................................................................................................................53

Index..........................................................................................................................................................54

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Ion Source Overview

1

The Turbo V

TM

DuoSpray ion source housing.

TM

ion source enables the use of either the TurboIonSpray

®

or APCI probe in the same

Use the ion source for either electrospray ionization (ESI), with the TurboIonSpray probe, or atmospheric pressure chemical ionization (APCI), with the APCI probe. Applications for the ion source include qualitative method development and qualitative and quantitative analysis.

The DuoSpray ion source with the optional calibrant delivery system (CDS) can be used to introduce calibration solution for automated mass calibration of the mass spectrometer. The CDS makes sure that the mass accuracy of the system is maintained throughout batch acquisition. For more information, refer to the Calibrant Delivery

System Operator Guide.

WARNING! Radiation Hazard, Biohazard, or Toxic Chemical Hazard: Use the ion source only if you have knowledge of and training in the proper use, containment, and evacuation of toxic or injurious materials used with the ion source. Discontinue use of the ion source if the window is cracked or damaged and contact an AB SCIEX Field Service Employee. Any toxic or injurious materials introduced into the equipment will be present in the ion source and exhaust output. Dispose of sharps following established laboratory safety procedures.

WARNING! Electrical Shock Hazard: Avoid contact with the high voltages applied to the ion source during operation. Put the system into Standby mode before adjusting the sample tubing or other equipment near the ion source.

Related Documentation

The guides and tutorials for the mass spectrometer and the Analyst

®

TF software are installed automatically with the software and are available from the Start menu: All Programs > AB SCIEX > Analyst TF. A complete list of the available documentation can be found in the Help. To view the software Help, press F1.

Technical Support

AB SCIEX and its representatives maintain a staff of fully-trained service and technical specialists located throughout the world. They can answer questions about the system or any technical issues that might arise. For more information, visit the Web site at www.absciex.com

.

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Ion Source Overview

Ion Source Components

Figure 1-1

shows the parts of the ion source.

Figure 1-1 Ion Source Components

Item

1

2

3

Description

Electrode adjustment cap

Corona discharge needle adjustment knob

APCI probe

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Ion Source Overview

8

9

6

7

Item

4

5

Description

Micrometer for the APCI probe used to position the probe on the vertical axis for ion source sensitivity adjustments

Corona discharge needle, which ionizes the trace species, or sample gas. Primary ions, which are formed as a result of the discharge, are converted by collisional processes to final ion-molecule reaction products.

Turbo heater

Micrometer for the TurboIonSpray

® sensitivity adjustments

probe used to position the horizontal axis for ion source

Micrometer for the TurboIonSpray probe used to position the vertical axis for ion source sensitivity adjustments

TurboIonSpray probe

Probes

The TurboIonSpray

®

probe and the APCI probe provide a range of capability for testing samples. Choose the probe and method most suitable for the compound in the sample stream flow.

The DuoSpray

TM

ion source is designed so that the probes cannot be installed interchangeably.

Table 1-1 Ion Source Specifications

Specification

Temperature

Liquid chromatography (LC)

Gas 1

Gas 2

Bath gas

TurboIonSpray

®

Probe

Probe temperature from

0°C to 750°C

APCI Probe

Probe temperature from

50°C to 750°C

Interfaces with any LC system

Nebulizer gas. UHP nitrogen

(99.999%) or an AB SCIEX recommended gas generator.

Heater gas. UHP nitrogen (99.999%) or an AB SCIEX recommended gas generator.

N/A

Nebulizer gas. UHP nitrogen (99.999%) or an AB SCIEX recommended gas generator.

UHP nitrogen (99.999%) or an AB SCIEX recommended gas generator.

If the optional CDS is used, then the user can switch between ionization modes by replumbing the sample and calibrant lines.

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Ion Source Overview

TurboIonSpray

®

Probe

The TurboIonSpray probe is ideally suited to LC/MS/MS analysis. It produces ions through ion evaporation. The sensitivity that is achieved with this technique is dependent on both flow rate and analyte. Because of better desolvation at higher flow rates, ionization efficiency increases with the increase in ion source temperature, which results in improved sensitivity. Compounds with extremely high polarity and low surface activity usually show the greatest sensitivity increase with an increase in source temperature. The TurboIonSpray

®

probe technique is mild enough to be used with labile compounds, such as peptides, proteins, and thermally labile pharmaceuticals.

When the heater is turned off, the TurboIonSpray probe functions as a conventional IonSpray

TM

ion source. It also functions with flow rates from 5 μL/min to 3000 μL/min and it vaporizes 100% aqueous to 100% organic solvents.

The TurboIonSpray probe consists of 0.012 inch outside diameter (o.d.) stainless steel tubing and is located centrally with a turbo heater placed at a 45 degree angle on the right side, when viewed from the front of the ion source.

Samples introduced through the TurboIonSpray probe are ionized within the tubing, by the application of high voltage (IonSpray voltage). Then they are nebulized by a jet of hot, dry, ultrahigh purity (UHP) nitrogen gas from the turbo heaters, creating a mist of small, highly-charged droplets. The combination of the IonSpray effluent and the heated dry gas from the turbo sprayer is projected at a 90 degree angle to the ion path. Refer to

Principles of Operation—Ion Source on page 39

.

Figure 1-2 Parts of the TurboIonSpray Probe

Item

1

2

3

Description

Electrode adjustment nut (black collar) that adjusts the extension of the electrode tip

Bronze retaining ring that fastens the probe to the probe tower on the ion source housing

Electrode tip through with samples are sprayed into the sample inlet area of the ion source

APCI Probe

The APCI probe is suitable for:

• Ionization of compounds that do not readily form ions in solution. These are usually non-polar compounds.

• Creation of simple APCI spectra for LC/MS/MS experiments.

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Ion Source Overview

• High-throughput analyses of complex and dirty samples. It is less sensitive to ion suppression effects.

• Rapid sample introduction by flow injection with or without an LC column.

The APCI probe can accept the entire effluent, without splitting, at flow rates from 50 μL/min to 3000 μL/min

(through a wide-bore column). It can vaporize volatile and labile compounds with minimal thermal decomposition.

The rapid desolvation and vaporization of the droplets and entrained analyte minimizes thermal decomposition and preserves molecular identity for ionization by the corona discharge needle. Buffers are readily tolerated by the ion source without significant contamination and the flash vaporization of the sprayed effluent allows up to

100% water to be used without difficulty.

The APCI probe consists of 100 μm inside diameter (i.d.) (0.004 inch) stainless steel tubing surrounded by a flow of nebulizer gas (Gas 2). The liquid sample stream is pumped through the sprayer, where it is nebulized in a ceramic tube containing a heater. The inside wall of the ceramic tube can be maintained at a temperature range of 100°C to 750°C and is monitored by the sensor embedded in the heater.

A high-velocity jet of nebulizer gas flows around the electrode tip to disperse the sample as a mist of fine particles.

It moves through the ceramic vaporization heater into the reaction region of the ion source and then past the corona discharge needle where the sample molecules are ionized as they pass through the ion source housing.

Refer to

Principles of Operation—Ion Source on page 39

.

Figure 1-3 Parts of the APCI Probe

Item

1

2

3

Description

Electrode adjustment nut (black collar) that adjusts the extension of the electrode tip

Bronze retaining ring that fastens the probe to the probe tower on the ion source housing

Electrode tip through which samples are sprayed into the sample inlet area of the ion source

Gas and Electrical Connections

Gas and high-voltage electrical connections are provided through the front plate of the interface and connect internally through the ion source housing. When the ion source is installed on the mass spectrometer, all of the electrical and gas connections are complete.

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Ion Source Overview

For more information about the function of Gas 1 and Gas 2 in the ion source, refer to

Source Parameters and Voltages on page 46

. Bath gas performs these functions:

• It prevents back streaming in to the tower part of the source, thus preventing accumulation of contaminants.

• It supplements the heater gas, providing a stream of gas that feeds into the entrainment region of the nebulizer gas expansion, thus reducing circulation and improving the spray columination.

• It helps to cool the tower region during high heat operation (such as in APCI mode).

Ion Source Sense Circuit

An ion source sense circuit disables the high-voltage power supply for the mass spectrometer and the source exhaust system if:

• The ion source housing is not installed or is improperly installed.

• A probe is not installed.

• The mass spectrometer senses a gas fault.

Source Exhaust System

WARNING! Radiation Hazard, Biohazard, or Toxic Chemical Hazard: Use the ion source only if you have knowledge of and training in the proper use, containment, and evacuation of toxic or injurious materials used with the ion source. Discontinue use of the ion source if the window is cracked or damaged and contact an AB SCIEX Field Service Employee. Any toxic or injurious materials introduced into the equipment will be present in the ion source and exhaust output. Dispose of sharps following established laboratory safety procedures.

WARNING! Radiation Hazard, Biohazard, or Toxic Chemical Hazard: Be sure to use the source exhaust system to safely remove sample vapor exhaust from the laboratory environment. For requirements for the source exhaust system, refer to the Site Planning Guide.

WARNING! Radiation Hazard, Biohazard, or Toxic Chemical Hazard: Vent the source exhaust system to either an external fume hood or an external vent to prevent hazardous vapors from being released into the laboratory environment.

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Ion Source Overview

WARNING! Fire Hazard: Do not direct more than 3 mL/min of solvent in to the ion source. Exceeding the maximum flow rate can cause solvent to accumulate in the ion source. Make sure that the source exhaust system is working, to prevent flammable vapor from accumulating in the ion source.

All ion sources produce both sample and solvent vapors. These vapors are a potential hazard to the laboratory environment. The source exhaust system is designed to safely remove and allow for the appropriate handling of the sample and solvent vapors. When the ion source is installed, the mass spectrometer will not operate unless the source exhaust system is operating.

A vacuum switch mounted in the source exhaust circuit measures the vacuum in the source. If the vacuum in the source rises above the set point while the probes are installed, the system goes into an exhaust fault (not ready) state.

An active exhaust system removes ion source exhaust (gases, solvent, sample vapor) through a drain port without introducing chemical noise. The drain port connects through a drain chamber and a source exhaust pump to a drain bottle, and from there to a customer-supplied exhaust ventilation system. For more information on the requirements for the source exhaust system, refer to the Site Planning Guide.

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Ion Source Installation

2

WARNING! Electrical Shock Hazard: Install the ion source on the mass spectrometer as the last step in this procedure. High voltage is present when the ion source is installed on the equipment.

The ion source is connected to the vacuum interface and is held in position by two source latches. The interior of the ion source is visible through the tempered glass windows on the side and end of the ion source.

When the ion source is installed, the software recognizes the ion source and displays the ion source identification.

Required Materials

• Ion source housing assembly

• TurboIonSpray

®

probe

• (Optional) APCI probe

• Ion source consumables kit

Prepare for Installation

WARNING! Radiation Hazard, Biohazard, or Toxic Chemical Hazard: Make sure that the electrode protrudes beyond the tip of the probe, to prevent hazardous vapors from escaping from the source. The electrode must not be recessed within the probe.

WARNING! Puncture Hazard: Be careful when handling the electrode tube. The tip of the electrode tube is extremely sharp.

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Ion Source Installation

Tip! Do not discard the empty package. Use it to store the ion source when it is not in use.

• Adjust the black electrode adjustment cap on the probe to move the electrode tip inside the electrode tube.

Install the Probes

The probes are not preinstalled in the ion source. Install the probes in the ion source housing before you install the ion source. Make sure that you insert each probe in the correct tower. The probes cannot be used interchangeably.

The probes can be inserted and removed separately as required. Refer to

Remove the Ion Source on page

28

.

If both probes are not properly installed in the ion source housing, then the Analyst

®

TF software reports that the ion source is not installed. High-voltage power for the mass spectrometer and source exhaust system are both turned off, and the Source/Gas tab in the Analyst TF software does not show voltage or temperature.

WARNING! Electrical Shock Hazard: Make sure that the ion source is completely disconnected from the mass spectrometer before proceeding.

WARNING! Electric Shock Hazard: Install the probe in the ion source before you install the ion source on the mass spectrometer.

CAUTION: Potential System Damage: Do not let the protruding electrode tip or the corona discharge needle touch any part of the ion source housing, to avoid damaging the probe.

1. Insert the APCI probe into the tower that is on the left side of the ion source when the glass window is facing you, inserting the raised plastic post into the groove on the probe.

2. Gently push down on the probe until the contacts engage with those in the tower.

3. Turn the bronze retaining ring over the probe, push it down to engage its threads with the threads on the tower, and then tighten the ring.

4. Insert the TurboIonSpray

®

probe into the tower on the top of the ion source, inserting the raised plastic post into the groove on the probe.

5. Gently push down on the probe until the contacts engage with those in the tower.

6. Turn the bronze retaining ring over the probe, push it down to engage its threads with the threads on the tower, and then tighten the ring until it is finger-tight

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Ion Source Installation

Connect the Ion Source Tubing

1. Insert a 30 cm (1 ft) piece of red PEEK tubing into the sample tubing nut.

2. Install the sample tubing nut on the fitting at the top of the probe.

3. Tighten the sample tubing nut until it is finger-tight.

Install the Ion Source on the Mass Spectrometer

WARNING! Electric Shock Hazard: Install the probe in the ion source before you install the ion source on the mass spectrometer.

Tip! Use the correct orifice plate for the system for optimal performance. Do not use an orifice plate for another system. The model number for the system is etched into the orifice plate.

If the ion source probe is not properly installed, then the high-voltage power supply is not available.

1. Make sure that the source latches on either side of the ion source are pointing up in the 12 o’clock position.

Refer to

Ion Source Components on page 6

.

2. Align the ion source with the vacuum interface, making sure that the latches on the ion source are aligned with the sockets in the vacuum interface.

3. Push the ion source gently against the vacuum interface and then rotate the ion source latches downwards to lock the ion source into place.

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Figure 2-1 Ion Source Latch

Ion Source Installation

The mass spectrometer recognizes the ion source and shows the ion source identification in the Analyst

®

TF software.

4. Connect the tubing from the sample supply device to the grounding union on the ion source.

Plumb the Ion Source for Sample Introduction with the TurboIonSpray

®

Probe

WARNING! Radiation Hazard, Biohazard, or Toxic Chemical Hazard: Make sure that the sample tubing nut is tightened properly before operating this equipment, to prevent leakage.

WARNING! Electrical Shock Hazard: Do not bypass the grounding union connection.

The grounding union provides grounding between the mass spectrometer and the sample introduction device.

1. Insert a 30 cm piece of red PEEK tubing into the sample tubing nut at the top of the TurboIonSpray probe.

2. Install the sample tubing nut on the fitting at the top of the TurboIonSpray probe.

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Ion Source Installation

3. Tighten the sample tubing nut until it is finger-tight.

4. Connect the other end of the red PEEK tubing to the grounding union.

5. Connect red PEEK tubing from the sample supply device to the grounding union on the ion source.

Plumb the Ion Source for Sample Introduction with the APCI Probe

WARNING! Radiation Hazard, Biohazard, or Toxic Chemical Hazard: Make sure that the sample tubing nut is tightened properly before operating this equipment, to prevent leakage.

WARNING! Electrical Shock Hazard: Do not bypass the grounding union connection.

The grounding union provides grounding between the mass spectrometer and the sample introduction device.

1. Insert a 30 cm piece of red PEEK tubing into the sample tubing nut at the top of the APCI probe.

2. Install the sample tubing nut on the fitting at the top of the APCI probe.

3. Tighten the sample tubing nut until it is finger-tight.

4. Connect the other end of the red PEEK tubing to the grounding union.

5. Connect red PEEK tubing from the sample supply device to the grounding union on the ion source.

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Ion Source Optimization

3

WARNING! Radiation Hazard, Biohazard, or Toxic Chemical Hazard: Use the ion source only if you have knowledge of and training in the proper use, containment, and evacuation of toxic or injurious materials used with the ion source. Discontinue use of the ion source if the window is cracked or damaged and contact an AB SCIEX Field Service Employee. Any toxic or injurious materials introduced into the equipment will be present in the ion source and exhaust output. Dispose of sharps following established laboratory safety procedures.

WARNING! Fire Hazard: Do not direct more than 3 mL/min of solvent in to the ion source. Exceeding the maximum flow rate can cause solvent to accumulate in the ion source. Make sure that the source exhaust system is working, to prevent flammable vapor from accumulating in the ion source.

Optimize the ion source whenever the analyte, flow rate, or mobile phase composition changes.

Several parameters affect the performance of the source. Optimize the performance while injecting a known compound and monitoring the signal of the known ion. Adjust the micrometer, gas, and voltage parameters to maximize the signal-to-noise ratio and signal stability.

If the optional CDS is used, then follow the instructions in the CDS User Guide to optimize the ion source.

Sample Introduction

Method

The liquid sample stream is delivered to the ion source by an LC pump or by a syringe pump. If it is delivered by an LC pump, then the sample can be injected directly into the mobile phase using FIA or tee infusion, or through a separation column using a loop injector or autosampler. If it is introduced by a syringe pump, then the sample is injected directly into the ion source. Infusion optimization can be used for ion path optimization and MS/MS fragment selection.

Flow Rate

Sample flow rates are determined by the chromatography system or by the volume of the sample available.

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Ion Source Optimization

Sample Inlet Requirements

• Use appropriate analytical procedures and practices to minimize external dead volumes. The sample inlet transfers the liquid sample to the ion source inlet without loss and with minimal dead volume.

• Prefilter samples so that the capillary tubing in the sample inlets is not blocked by particles, precipitated samples, or salts.

• Make sure that all connections are tight enough to prevent leaks. Do not over tighten.

TurboIonSpray

®

Probe Optimization

WARNING! Radiation Hazard, Biohazard, or Toxic Chemical Hazard: Make sure that the mass spectrometer is properly vented and that good general laboratory ventilation is provided. Adequate laboratory ventilation is required to control solvent and sample emissions and to provide for safe operation of the mass spectrometer.

CAUTION: Potential System Damage: If the LC system connected to the mass spectrometer is not controlled by the Analyst

®

TF software, then do not leave the mass spectrometer unattended while in operation. The LC system can flood the ion source when the mass spectrometer goes into Standby mode.

Several parameters affect the performance of the TurboIonSpray probe. Optimize the performance while injecting a known compound and monitoring the signal of the known ion. Adjust the parameters to maximize the

signal-to-noise ratio and signal stability. Refer to

TurboIonSpray

®

Probe Parameters on page 46

.

Note: If the IonSpray Voltage Floating (ISVF) is too high, then a corona discharge can occur. It is visible as a blue glow at the tip of the TurboIonSpray

®

probe. A corona discharge results in decreased sensitivity and stability of the ion signal.

Note: The IonSpray Voltage Floating (ISVF) is always applied to both the TurboIonSpray

®

probe and the APCI probe simultaneously, and the Temperature (TEM) is always applied to both the turbo and APCI heaters simultaneously.

Note: To keep the system clean and at its optimum performance, adjust the probe position when changing the flow rate.

Tip! It is easier to optimize signal and signal-to-noise with flow injection analysis or on-column injections.

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Ion Source Optimization

Flow Rate and Temperature

The quantity and type of sample affects the optimal TurboIonSpray

®

probe temperature. At higher flow rates, the optimal temperature increases. A more significant factor is the composition of the solvent. As the organic content of the solvent increases, the optimal probe temperature should decrease.

The TurboIonSpray probe is normally used with sample flow rates of 40 µL/min to 1000 µL/min. The heat is used to increase the rate of evaporation which improves ionization efficiency, resulting in increased sensitivity. Extremely low flow rates of high organic solvent usually do not require increased temperatures.

Set Up the System

1. Configure the HPLC pump to deliver the mobile phase at the required flow rate. Refer to

Source Parameters and Voltages on page 46

.

2. Connect the grounding union on the ion source to a pump, through an injector equipped with a 5 µL loop, or to an autosampler.

3. If using an autosampler, then configure the autosampler to perform multiple injections.

Run the Method

1. Start the Analyst

®

TF software.

2. In the Navigation bar, under Tune and Calibrate mode, double-click Manual Tuning.

3. Open a previously optimized method or create a method based on the compounds.

4. If the ion source has been allowed to cool, then do the following.

a. Set the Temperature (TEM) parameter to 450.

b. Let the ion source warm up for 30 minutes.

The 30-minute warm-up stage prevents solvent vapors from condensing in the cold probe.

5. Start acquisition.

6. Start the sample flow and sample injection.

Set the Starting Conditions

1. On the Source/Gas tab in the Tune Method Editor, type a starting value for Ion Source Gas 1 (GS1).

For LC pumps, use a value between 40 and 60 for GS1.

2. Type a starting value for Ion Source Gas 2 (GS2).

For LC pumps, use a value between 30 and 50 for GS2.

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Ion Source Optimization

Note: Gas 2 is used with higher flow rates typical with an LC system and in conjunction with increased temperature.

3. Type 5500 in the IonSpray Voltage Floating (ISVF) field.

4. Type 25 in the Curtain Gas (CUR) field.

Optimize the TurboIonSpray

®

Probe Position

1. Look through the window of the ion source housing to view the position of the probe.

2. Use the previous horizontal and vertical micrometer settings or set them to 5 as a starting position.

3. Use FIA or a tee infusion to inject sample at a high flow rate.

4. Monitor the signal in the software.

5. Use the horizontal micrometer to adjust the probe position in small increments to achieve the best signal or signal-to-noise ratio.

The probe can optimize slightly to either side of the aperture.

Tip! It is easier to optimize signal and signal-to-noise with flow injection analysis or on-column injections.

6. Use the vertical micrometer to adjust the probe position in small increments to achieve the best signal or signal-to-noise ratio.

Note: The vertical position of the probe depends on flow rate. At lower flow rates, the probe should be closer to the aperture. At higher flow rates, the probe should be farther from the aperture.

WARNING! Radiation Hazard, Biohazard, or Toxic Chemical Hazard: Make sure that the electrode protrudes beyond the tip of the probe, to prevent hazardous vapors from escaping from the source. The electrode must not be recessed within the probe.

7. Adjust the black electrode adjustment cap on the probe to extend the electrode tip. Typically, the optimum extension of the electrode is 0.5 mm to 1.0 mm beyond the end of the probe.

After the probe is optimized, it needs only minor adjustment. If the probe is removed, or if the analyte, flow rate, or solvent composition change, repeat the optimizing procedure after installation.

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Ion Source Optimization

Tip! Direct the liquid spray from the TurboIonSpray probe away from the aperture in order to prevent contamination of the aperture, to prevent piercing of the Curtain Gas

TM

flow, which can create an unstable signal, and to prevent electrical shorting due to the presence of the liquid.

Optimize Source and Gas Parameters and Voltage

Optimize Gas 1 (nebulizer gas) for best signal stability and sensitivity. Gas 2 (heater gas) aids in the evaporation of solvent, which helps to increase the ionization of the sample.

Too high a temperature can cause premature vaporization of the solvent at the TurboIonSpray

®

probe tip, especially if the probe is too low, which will result in signal instability and a high chemical background noise. Similarly, a high heater gas flow could produce a noisy or unstable signal.

Use the lowest IonSpray

TM

source voltage possible without losing signal. Focus on signal-to-noise and not just signal. If the IonSpray source voltage is too high, then a corona discharge can occur. It is visible as a blue glow at the tip of the TurboIonSpray probe. This will result in decreased sensitivity and stability of the ion signal.

1. Adjust GS1 and GS2 in increments of 5 to achieve the best signal or signal-to-noise ratio.

Note: To prevent contamination, use the highest value for CUR possible without sacrificing sensitivity. Do not set CUR lower than 20. This helps to prevent penetration of the Curtain Gas flow, which can produce a noisy signal, prevent contamination of the aperture and increase the overall signal-to-noise ratio.

2. Increase the value in the CUR field until the signal begins to decrease.

3. Adjust ISVF in increments of 500 V to maximize signal-to-noise.

Note: If the IonSpray Voltage Floating (ISVF) is too high, then a corona discharge can occur. It is

® visible as a blue glow at the tip of the TurboIonSpray probe. A corona discharge results in decreased sensitivity and stability of the ion signal.

Optimize the Turbo Heater Temperature

The optimal heater temperature is compound-dependent, flow rate-dependent, and mobile phase composition-dependent. The higher the flow rate and the higher the aqueous composition, the higher the optimized temperature.

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Ion Source Optimization

When optimizing the source temperature, make sure that the ion source equilibrates to the new temperature setting.

• Adjust the TEM value in increments of 50°C to 100°C to achieve the best signal or signal-to-noise ratio.

APCI Probe Optimization

WARNING! Radiation Hazard, Biohazard, or Toxic Chemical Hazard: Make sure that the mass spectrometer is properly vented and that good general laboratory ventilation is provided. Adequate laboratory ventilation is required to control solvent and sample emissions and to provide for safe operation of the mass spectrometer.

CAUTION: Potential System Damage: If the LC system connected to the mass spectrometer is not controlled by the Analyst

®

TF software, then do not leave the mass spectrometer unattended while in operation. The LC system can flood the ion source when the mass spectrometer goes into Standby mode.

Refer to

APCI Probe Parameters on page 47

.

CAUTION: It is easier to optimize signal and signal-to-noise with flow injection analysis or on-column injections.

Note: The IonSpray Voltage Floating (ISVF) is always applied to both the TurboIonSpray

®

probe and the APCI probe simultaneously, and the Temperature (TEM) is always applied to both the turbo and APCI heaters simultaneously.

Set Up the System

1. Configure the HPLC pump to deliver the mobile phase at the required flow rate. Refer to

Source Parameters and Voltages on page 46

.

2. Connect the grounding union on the ion source to a pump, through an injector equipped with a 5 µL loop, or to an autosampler.

3. If using an autosampler, then configure the autosampler to perform multiple injections.

Run the Method

1. Start the Analyst

®

TF software.

2. In the Navigation bar, under Tune and Calibrate mode, double-click Manual Tuning.

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Ion Source Optimization

3. Open a previously optimized method or create a method based on the compounds.

4. If the ion source has been allowed to cool, then do the following.

a. Set the Temperature (TEM) parameter to 450.

b. Let the ion source warm up for 30 minutes.

The 30-minute warm-up stage prevents solvent vapors from condensing in the cold probe.

5. Start acquisition.

6. Start the sample flow and sample injection.

Set the Starting Conditions

Note: The value for the GS1 parameter, which is used by the TurboIonSpray

®

probe, might influence performance of the APCI probe. Adjust the GS1 parameter value to achieve optimal performance.

Note: Gas 2 is used as a nebulizer gas for the APCI probe.

1. Type 20 in the Ion Source Gas 2 (GS2) field.

2. Type 25 in the Curtain Gas (CUR) field.

3. Type 5500 in the IonSpray Voltage Floating (ISVF) field.

Optimize Gas 2 and Curtain Gas

TM

Flow

1. Adjust GS2 in increments of five to achieve the best signal or signal-to-noise ratio.

Note: To prevent contamination, use the highest value for CUR possible without sacrificing sensitivity. Do not set CUR lower than 20. This helps to prevent penetration of the Curtain Gas flow, which can produce a noisy signal, prevent contamination of the aperture and increase the overall signal-to-noise ratio.

2. Increase CUR until the signal starts to decrease.

Adjust the Position of the Corona Discharge Needle

WARNING! Electrical Shock Hazard: Follow this procedure to avoid contact with the high voltages applied to the corona discharge needle, curtain plate, and turbo heaters.

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Ion Source Optimization

When using the APCI probe, make sure that the corona discharge needle is pointing toward the aperture.

Required Materials

• Insulated flat-bladed screwdriver

1. Use an insulated flat-bladed screwdriver to rotate the corona discharge needle adjustment screw on the top of the needle.

2. Look through the glass window to make sure that the needle is aligned with the tip facing the aperture.

3. Save the optimized method as a new method.

Optimize the APCI Probe Position

Make sure that the curtain plate aperture remains clear of solvent or solvent droplets at all times.

The position of the sprayer nozzle affects sensitivity and signal stability. Adjust probe sensitivity in small increments only. At lower flow rates, position the probe closer to the aperture. For higher flow rates, position the probe farther away from the aperture.

Figure 3-1 Sprayer Nozzle Position

Item

1

2

3

Description

Corona discharge needle

Curtain plate

APCI probe

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Ion Source Optimization

1. Use the previous setting or set it to 5 mm as an initial starting position.

Note: To avoid reducing the performance of the mass spectrometer, do not spray directly into the aperture.

2. Use FIA or a tee infusion to inject sample at a high flow rate.

3. Monitor the signal in the software.

4. Use the vertical micrometer to adjust the probe in small increments to achieve the best signal or signal-to-noise ratio.

After the probe is optimized, it needs only minor adjustment. If the probe is removed, or if the analyte, flow rate, or solvent composition changes, repeat the optimizing procedure after installation.

Optimize the IonSpray

Voltage Floating

Note: If the IonSpray Voltage Floating (ISVF) is too high, then a corona discharge can occur. It is visible as a blue glow at the tip of the TurboIonSpray

®

probe. A corona discharge results in decreased sensitivity and stability of the ion signal.

• In positive mode, start at a value of 5500, and decrease in steps of 100 V to 500 V; in negative mode, start at a value of –4500, and increase in steps of 100 V to 500 V. Continue adjusting to achieve the best signal or signal-to-noise ratio.

This parameter usually optimizes around 5500 V in positive mode. If you observe no changes in signal with increasing ISVF, then leave the ISVF at the lowest setting that provides the best signal or signal-to-noise ratio.

Optimize the APCI Probe Temperature

WARNING! Radiation Hazard, Biohazard, or Toxic Chemical Hazard: Make sure that the mass spectrometer is properly vented and that good general laboratory ventilation is provided. Adequate laboratory ventilation is required to control solvent and sample emissions and to provide for safe operation of the mass spectrometer.

The quantity and type of solvent affects the optimal APCI probe temperature. At higher flow rates, the optimal temperature increases.

• Adjust the TEM value in increments of 50°C to 100°C to achieve the best signal or signal-to-noise ratio.

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Ion Source Maintenance

4

The following warnings apply to all maintenance procedures in this chapter.

WARNING! Hot Surface Hazard: Let the ion source cool for at least 30 minutes before starting any maintenance procedures. Surfaces of the ion source and the vacuum interface components become hot during operation.

WARNING! Fire and Toxic Chemical Hazard: Keep flammable solvents away from flame and sparks and use them only in vented chemical fume hoods or safety cabinets.

WARNING! Toxic Chemical Hazard: Wear personal protective equipment, including a laboratory coat, gloves, and safety glasses to avoid skin or eye exposure to solvents.

WARNING! Radiation Hazard, Biohazard, or Toxic Chemical Hazard: In the event of a chemical spill, review product Safety Data Sheets for specific instructions. Stop the spill or leak only if it is safe to do so. Use appropriate personal protective equipment and absorbent wipes to contain the spill and dispose of it following local regulations.

WARNING! Electrical Shock Hazard: Avoid contact with the high voltages applied to the ion source during operation. Put the system into Standby mode before adjusting the sample tubing or other equipment near the ion source.

This section contains general maintenance procedures for the ion source. To determine how often to clean the ion source or perform preventive maintenance, consider the following:

• Compounds tested

• Cleanliness of the preparation methods

• Amount of time an idle probe contains a sample

• Overall system run time

These factors can cause changes in ion source performance, indicating that maintenance is required.

Make sure that the mounted ion source is fully sealed to the mass spectrometer with no evidence of gas leaks.

Perform general maintenance inspections to be sure of safe operation of the system. Clean the ion source components regularly to keep the ion source in good working condition.

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Ion Source Maintenance

CAUTION: Potential System Damage: Use only the recommended cleaning method to avoid damaging the equipment.

Required Materials

• 1/4 inch open-ended wrench

• 9/64 inch hex key (L-shaped key supplied)

• 5 mm hex key

• 2.5 mm hex key

• Phillips screwdriver

• Flat-bladed screwdriver

• MS-grade methanol

• HPLC-grade deionized water

• Safety glasses

• Breathing mask and filter

• Powder-free gloves (neoprene is recommended)

• Lab coat

Clean the Probes

WARNING! Hot Surface Hazard: Let the ion source cool for at least 30 minutes before starting any maintenance procedures. Surfaces of the ion source and the vacuum interface components become hot during operation.

Flush the ion source periodically, regardless of the type of compounds sampled. Do this by setting up a method in the software specifically for performing a flushing operation.

1. Switch to a mobile phase that is 1:1 water:acetonitrile or 1:1 water:methanol.

2. Adjust the position of the probes so that they are as far from the orifice as possible.

3. In the software do the following.

a. Set Temperature (TEM) between 500 and 600.

b. Set Ion Source Gas (GS1) and Ion Source Gas 2 (GS2) to at least 40.

c. Set Curtain Gas (CUR) to the highest setting possible without loss of signal.

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Ion Source Maintenance

d. Wait until the TEM setpoint is reached.

4. Direct the flow of mobile phase through the tubing and each probe at 1 mL/min for 10 minutes to 15 minutes.

5. Make sure that both probes and sample tubing are flushed thoroughly.

Remove the Ion Source

WARNING! Hot Surface Hazard: Let the ion source cool for at least 30 minutes before starting any maintenance procedures. Surfaces of the ion source and the vacuum interface components become hot during operation.

Note: An additional 5.3 L/min of nitrogen flows when the mass spectrometer is off or the ion source is removed from the system. To minimize nitrogen gas consumption and to keep the mass spectrometer clean when it is not in use, leave the ion source installed on the mass spectrometer and leave the system on.

The ion source can be removed quickly and easily, without tools. Always remove the ion source from the mass spectrometer before performing any maintenance on the ion source or exchanging the probes.

1. Stop any ongoing scans.

2. Shut down the sample stream.

3. Type 0 in the Temperature (TEM) field, if the heaters are in use.

4. Disconnect the sample tubing from the grounding union.

5. Turn the two source latches upward to the 12 o'clock position to release the ion source.

6. Pull the ion source gently away from the vacuum interface.

7. Put the ion source on a clean, secure surface.

Remove the Probe

Prerequisite Procedures

Remove the Ion Source on page 28

WARNING! Hot Surface Hazard: Let the ion source cool for at least 30 minutes before starting any maintenance procedures. Surfaces of the ion source and the vacuum interface components become hot during operation.

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Ion Source Maintenance

WARNING! Electrical Shock Hazard: Make sure that the ion source is completely disconnected from the mass spectrometer before proceeding.

The probe can be removed quickly and easily, without tools. Always remove the ion source from the mass spectrometer before changing probes or performing maintenance on the probe.

When replacing the probes, insert them into the correct tower. The probes cannot be used interchangeably. Refer

to

Install the Probes on page 13

.

1. Loosen the sample tubing nut and then disconnect the sample tubing from the probe.

2. Loosen the brass retaining ring that fastens the probe to the ion source housing.

3. Gently pull the probe straight up out of the tower.

Note: Do not let the tip of the probe touch anything while it is being removed or stored.

4. Put the probe on a secure, clean surface.

Clean the Electrode Tube

Prerequisite Procedures

Remove the Ion Source on page 28

Remove the Probe on page 28

WARNING! Electrical Shock Hazard: Make sure that the ion source is completely disconnected from the mass spectrometer before proceeding.

WARNING! Puncture Hazard: Be careful when handling the electrode tube. The tip of the electrode tube is extremely sharp.

WARNING! Hot Surface Hazard: Let the ion source cool for at least 30 minutes before starting any maintenance procedures. Surfaces of the ion source and the vacuum interface components become hot during operation.

The probe contains an electrode tube. Clean the electrode tube periodically, or when there is a decrease in performance.

This procedure applies to both probes. Use this procedure to remove the electrode tube for cleaning. If the electrode tube cannot be cleaned, then use this procedure to replace it.

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Ion Source Maintenance

1. Remove the electrode adjustment nut.

2. Holding the probe with the tip pointing downwards, so that the spring remains inside the probe, pull the PEEK union and the attached electrode tube from the probe.

Figure 4-1 Probe, Expanded View

4

5

6

Item Description

1

2

3

Electrode adjustment nut

1/4 inch retaining nut

Spring

7

8

Bronze retaining ring

Sprayer tube

Electrode Tip

Electrode tube

PEEK union

3. Use the 1/4 inch open-ended wrench to remove the retaining nut that holds the electrode tube in the PEEK union.

4. Remove the electrode tube from the retaining nut.

5. Clean the electrode tube with a 1:1 methanol:water solution by soaking the tube in an ultrasonic bath.

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Ion Source Maintenance

Assemble the Probe Components

WARNING! Puncture Hazard: Be careful when handling the electrode tube. The tip of the electrode tube is extremely sharp.

When the electrode tube is cleaned, or replaced with a new part, assemble the probe components.

1. Make sure that the spring is still inside the probe and then tighten the electrode adjustment nut.

2. Align the electrode tube with the narrow opening in the sprayer tube and then insert the PEEK union and attached electrode tube into the probe. Be careful not to bend the electrode tube.

3. Insert the probe into the tower, taking care not to allow the tip of the probe to touch any part of the ion source housing.

4. Push down the brass retaining ring to engage its thread with the thread on the ion source housing and then tighten the ring.

5. Insert a 30 cm piece of red PEEK tubing into the sample tubing nut.

6. Install the sample tubing nut in the fitting at the top of the probe, and then tighten the sample tubing nut until it is finger-tight.

7. Install the ion source on the mass spectrometer. Refer to

Ion Source Installation on page 12

.

8. Adjust the electrode tip extension. Refer to

Adjust the Electrode Tip Extension on page 31

.

Adjust the Electrode Tip Extension

WARNING! Radiation Hazard, Biohazard, or Toxic Chemical Hazard: Make sure that the electrode protrudes beyond the tip of the probe, to prevent hazardous vapors from escaping from the source. The electrode must not be recessed within the probe.

WARNING! Puncture Hazard: Be careful when handling the electrode tube. The tip of the electrode tube is extremely sharp.

Adjust the electrode tip extension for best performance. The optimal setting is compound-dependent. The distance that the electrode tip extends affects the shape of the spray cone, and the shape of the spray cone affects mass spectrometer sensitivity.

• Adjust the black electrode adjustment cap on the top of the probe to extend or retract the electrode tip. The electrode tip should extend between 0.5 mm and 1.0 mm from the end of the probe.

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Ion Source Maintenance

Figure 4-2 Electrode Tip Extension Adjustment

Item

1

2

Description

Probe

Electrode

Replace the Corona Discharge Needle Tip

Replace the tip of the corona discharge needle if it becomes corroded.

WARNING! Hot Surface Hazard: Let the ion source cool for at least 30 minutes before starting any maintenance procedures. Surfaces of the ion source and the vacuum interface components become hot during operation.

WARNING! Electrical Shock Hazard: Make sure that the ion source is completely disconnected from the mass spectrometer before proceeding.

WARNING! Puncture Hazard: Handle the needle with care. The tip of the needle is extremely sharp.

1. Remove the ion source from the mass spectrometer. Refer to

Remove the Ion Source on page 28

.

2. Rotate the ion source so that the open side is toward you.

3. Press down on the corona discharge needle adjustment knob on the top of the tower. The corona discharge needle extends.

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Ion Source Maintenance

4. Holding the corona discharge needle tip between the thumb and forefinger of one hand and the corona discharge needle with the other hand, rotate the corona discharge needle tip counter-clockwise to loosen and gently remove the tip.

Figure 4-3 Corona Discharge Needle Tip at Back of Ion Source

5. Holding a new tip between the thumb and forefinger of one hand and the corona discharge needle with the other hand, rotate the corona discharge needle tip clockwise to install the tip.

6. Install the ion source on the mass spectrometer. Refer to

Ion Source Installation on page 12

.

Replace the Corona Discharge Needle

Prerequisite Procedures

Remove the Ion Source on page 28

Remove the Probe on page 28

WARNING! Hot Surface Hazard: Let the ion source cool for at least 30 minutes before starting any maintenance procedures. Surfaces of the ion source and the vacuum interface components become hot during operation.

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Ion Source Maintenance

WARNING! Electrical Shock Hazard: Make sure that the ion source is completely disconnected from the mass spectrometer before proceeding.

WARNING! Puncture Hazard: Handle the needle with care. The tip of the needle is extremely sharp.

The corona discharge needle tip may become so corroded that it must be cut off the needle. If this occurs, replace the entire corona discharge needle.

1. Rotate the ion source so the open side is accessible.

Figure 4-4 Corona Discharge Needle

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Ion Source Maintenance

1

2

3

Item Description

Exhaust chimney

Ceramic sleeve

Corona discharge needle tip

2. Gently pull the corona discharge needle down through the exhaust chimney to remove it.

3. Insert the new needle through the exhaust chimney into the ceramic sleeve as far as it will go.

4. Holding a new tip between the thumb and forefinger of one hand and the corona discharge needle with the other hand, rotate the corona discharge needle tip clockwise to install the tip.

5. Insert the probe and then install the ion source on the mass spectrometer. Refer to

Ion Source Installation on page 12

.

Replace the Sample Tubing

WARNING! Electrical Shock Hazard: Avoid contact with the high voltages applied to the ion source during operation. Put the system into Standby mode before adjusting the sample tubing or other equipment near the ion source.

Use the following procedure to replace the sample tubing if it has a blockage.

1. Stop the sample flow and make sure that any remaining gas has been removed through the source exhaust

system. Refer to

Remove the Ion Source on page 28

.

2. Disconnect the sample tubing from the probe and the union.

3. Replace the sample tubing with the same length of tubing used previously.

4. Install the ion source. Refer to

Ion Source Installation on page 12

.

5. Start the sample flow.

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Troubleshooting Tips

5

Symptom

The Analyst

®

TF software reports that the mass spectrometer is in Fault status.

Possible Cause

The probe is not installed.

Corrective Action

Install the probe. Refer to

Install the Probes on page 13

.

The probe is not connected securely.

Remove and replace the probe.

Tighten the probe connection bronze ring securely. Refer to

Remove the

Probe on page 28

and

Install the Probes on page 13

.

The heater does not work.

The spray is not uniform.

F3 fuse is blown.

The electrode is blocked.

Contact an FSE.

Clean or replace the electrode. Refer to

Clean the Electrode Tube on page 29

.

Sensitivity is poor.

During testing, the ion source fails to meet specifications.

The interface components (front end) are dirty.

Clean the interface components and reposition the ion source.

Solvent vapor or other unknown compounds are present in the analyzer region.

The mass spectrometer has not passed the installation tests.

Optimize the Curtain Gas

Refer to

Optimization on page 17

source.

Ion Source

flow.

.

Perform installation tests on the mass spectrometer with the default

The test solution was not prepared correctly.

• Confirm that the test solutions were prepared correctly.

• If the problem cannot be resolved, contact the FSE.

Background noise is high.

The Temperature (TEM) is too high.

Optimize the temperature.

The heater gas flow rate (GS2) is too high.

Optimize heater gas flow.

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Symptom

Ion source performance has degraded.

Troubleshooting Tips

Possible Cause

The ion source is contaminated.

The probe is not optimized.

The sample was not prepared correctly or the sample has degraded.

Corrective Action

• Clean or replace ion source

components. Refer to

Ion

Source Maintenance on page 26

.

• Condition the source and front end: a. Move the probes to the furthest position from the aperture (vertically and horizontally).

b. Make sure the interface heater is On.

c. Infuse or inject 1:1 methanol:water with a pump flow of 1 mL/min.

d. In the Analyst TF software, set TEM to 650, GS1 to 60, and GS2 to 60.

e. Set the Curtain Gas flow to 45 or 50.

f.

Run for a minimum of 2 hours or preferably overnight for best results.

Refer to

APCI Probe

Optimization on page 22

and

TurboIonSpray

®

Probe

Optimization on page 18

.

Confirm that the sample was prepared correctly.

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Troubleshooting Tips

Symptom

Arcing or sparks occur.

Possible Cause Corrective Action

The sample inlet fittings are leaking.

• Verify that the fittings are tight and replace fittings if leaks continue. Do not overtighten the fittings.

• Install and optimize an alternate ion source. If the problem persists, contact an FSE.

The position of the corona discharge needle is incorrect.

Turn the corona discharge needle toward the curtain plate, and away from the stream of heater gas. Refer to

Adjust the Position of the

Corona Discharge Needle on page 23

.

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Principles of Operation—Ion

Source

A

TurboIonSpray

®

Mode

The TurboIonSpray probe uses a turbo heater to blow hot, dry, UHP (ultrahigh purity) nitrogen.The heater is at a

45 degree angle to the probe, on the right side, when viewed from the front of the ion source. The combination of IonSpray

TM

effluent and the heated dry gas from the turbo heaters is projected at a 90-degree angle to the aperture in the curtain plate.

Only compounds that ionize in the liquid solvent can be generated as gas phase ions in the source. The efficiency and rate of ion generation depends on the solvation energies of the specific ions. Ions with lower solvation energies are more likely to evaporate than ions with higher solvation energies.

The interaction of the IonSpray and theturbo heater helps focus the stream and increases the rate of droplet evaporation, resulting in an increased ion signal. The heated gas increases the efficiency of ion evaporation, resulting in increased sensitivity and the ability to handle higher liquid sample flow rates.

A high-velocity flow of nebulizer gas shears droplets from the liquid sample stream in the IonSpray inlet. Using the variable high voltage applied to the sprayer, the ion source applies a net charge to each droplet. This charge aids in the droplet dispersion. Ions of a single polarity are preferentially drawn into the droplets by the high voltage as they are separated from the liquid stream. However, this separation is incomplete and each droplet contains many ions of both polarities. Ions of one polarity are predominant in each droplet, and the difference between the number of positively or negatively charged ions results in the net charge. Only the excess ions of the predominant polarity are available for ion evaporation, and only a fraction of these actually evaporate.

The polarity and concentration of excess ions depends on the magnitude and polarity of the high-voltage potential applied to the sprayer tip. For example, when a sample contains arginine in a water-acetonitrile solution and a positive potential is applied to the sprayer, the excess positive ions will be H+ and MH+ arginine.

The probe can generate multiply-charged ions from compounds that have multiple charge sites, such as peptides and oligonucleotides. This is useful when observing high-molecular-weight species where the multiple charges produce ions of a mass-to-charge (m/z) ratio within the mass range of the mass spectrometer. This allows routine molecular-weight determinations of compounds in the kiloDalton (kDa) range.

As shown in

Figure A-1

, each charged droplet contains solvent and both positive and negative ions, but with

ions of one predominant polarity. As a conducting medium, excess charges reside at the surface of the droplet.

As the solvent evaporates, the electrical field at the surface of the droplet increases due to the decreasing radius of the droplet.

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Principles of Operation—Ion Source

Figure A-1 Ion Evaporation

Item

1

2

3

4

Description

Droplet contains ions of both polarities with one polarity being predominant.

As the solvent evaporates, the electrical field increases and the ions move to the surface.

At some critical field value, ions are emitted from the droplets.

Nonvolatile residue remains as a dry particle.

If the droplet contains excess ions and enough solvent evaporates from the droplet, a critical field is reached at which ions are emitted from the surface. Eventually, all of the solvent will evaporate from the droplet, leaving a dry particle consisting of the nonvolatile components of the sample solution.

Because the solvation energies for most organic molecules are unknown, the sensitivities of any given organic ion to ion evaporation are difficult to predict. The importance of solvation energy is evident because surfactants that concentrate at the surface of a liquid can be detected very sensitively.

APCI Mode

The basis for past incompatibilities in linking liquid chromatography with mass spectrometry arose from difficulties converting relatively involatile molecules in solution in a liquid into a molecular gas without inducing excessive decomposition. The APCI probe process of gently nebulizing the sample into finely dispersed small droplets in a heated ceramic tube results in the rapid vaporization of the sample so that the sample molecules are not decomposed.

Figure A-2

shows the reaction flow of the APCI process for reactant positive ions (the proton hydrates,

H

3

O

+

[H

2

O] n

).

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Figure A-2 APCI Reaction Flow Diagram

Principles of Operation—Ion Source

The major primary ions N2

+

, O

2

+

, H

2

O

+

, and NO

+

are formed by the electron impact of corona created electrons on the major neutral components of air. Although NO

+

is normally not a major constituent of clean air, the concentration of this species in the source is enhanced due to neutral reactions initiated by the corona discharge.

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Principles of Operation—Ion Source

Samples that are introduced through the APCI probe are sprayed, with the aid of a nebulizer gas, into the heated ceramic tube. Within the tube, the finely dispersed droplets of sample and solvent undergo a rapid vaporization with minimal thermal decomposition. The gentle vaporization preserves the molecular identity of the sample.

The gaseous sample and solvent molecules pass into the ion source housing where the ionization by APCI is induced by a corona discharge needle connected to the end of the ceramic tube. The sample molecules are ionized by colliding with the reagent ions created by the ionization of mobile phase solvent molecules. As shown in

Figure

A-3

, the vaporized solvent molecules ionize to produce the reagent ions [X+H]+ in the positive mode and [X-H]– in the negative mode. It is these reagent ions that produce stable sample ions when they collide with the sample molecules.

Figure A-3 Atmospheric Pressure Chemical Ionization

Item

1

2

3

4

Description

Sample

Primary ions are created in the vicinity of the corona discharge needle

Ionization produces predominantly solvent ions

Reagent ions react with sample molecules forming clusters

5

6

Curtain plate

Interface x = solvent molecules; M=sample molecules

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Principles of Operation—Ion Source

The sample molecules are ionized through a process of proton transfer in the positive mode and by either electron transfer or proton transfer in the negative mode. The energy for the APCI ionization process is collision-dominated because of the relatively high atmospheric pressure of the API source.

For reverse phase applications, the reagent ions consist of protonated solvent molecules in the positive mode and solvated oxygen ions in the negative mode. With favorable thermodynamics, the addition of modifiers changes the reagent ion composition. For example, the addition of acetate buffers or modifiers can make the acetate ion

[CH

[NH

3

4

COO]

]

+

the primary reagent in the negative mode. Ammonium modifiers may make protonated ammonia

the primary reagent in the positive mode.

Through collisions, an equilibrium distribution of certain ions (for example, protonated water cluster ions) is maintained. The likelihood of premature fragmentation of the sample ions in the ion source is reduced because of the moderating influence of solvent clusters on the reagent ions and the relatively high gas pressure in the source. As a result, the ionization process yields primarily molecular product ions for mass analysis in the mass spectrometer.

APCI Ionization Region

Figure A-4

shows the general location of the ion-molecule reactor of the APCI probe. The slanted lines indicate a wall-less reactor. A self-starting corona discharge ion current in the microampere range is created as a result of the electric field between the discharge needle and the curtain plate. Primary ions, for example, N

2

+

and O

2

+

, are created by the loss of electrons that originate in the plasma in the immediate vicinity of the discharge needle tip. The energy of these electrons is moderated by a number of collisions with gas molecules before attaining an energy where their effective ionization cross-section allows them to ionize neutral molecules efficiently.

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Principles of Operation—Ion Source

Figure A-4 APCI Ionization Region

5

6

7

8

Item

3

4

1

2

Description

Discharge needle tip

Sample flow

Wall-less reactor

Curtain plate aperture

Curtain Gas

TM

supply

Orifice

Orifice plate

Ceramic tube

The primary ions, in turn, generate intermediate ions that lead to the formation of sample ions. Ions of the chosen polarity drift under the influence of the electric field in the direction of the curtain plate and through the gas curtain into the mass analyzer. The whole ion formation process is collision-dominated because of the relatively high atmospheric pressure of the APCI probe. Except in the immediate vicinity of the discharge needle tip, where the

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Operator Guide

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Principles of Operation—Ion Source

strength of the electric field is greatest, the energy imparted to an ion by the electric field is small in comparison with the thermal energy of the ion.

Through collisions, an equal distribution of certain ions (for example, protonated water cluster ions) is maintained.

Any excess energy that an ion may acquire in the ion-molecule reaction process is thermalized. Through collisional stabilization, many of the product ions are fixed, even though many subsequent collisions occur. The formation of both product ions and reactant ions is governed by equilibrium conditions at 760 torr (atmospheric) operating pressure.

The APCI probe functions as a wall-less reactor because the ions that pass from the source to the vacuum chamber and eventually to the detector never experience collisions with a wall—only collisions with other molecules. Ions are also formed outside the designated APCI source, but are not detected and are eventually neutralized by interacting with a wall surface.

The temperature of the probe is an important factor for APCI probe operation. To preserve the molecular identity, the temperature must be set high enough to ensure a rapid evaporation. At a sufficiently high operating temperature, the droplets are vaporized quickly so that organic molecules are desorbed from the droplets with minimal thermal degradation. If, however, the temperature is set too low, the evaporation process is slower and pyrolysis, or decomposition, may occur before vaporization is complete. Operating the APCI probe at temperatures above the optimal temperature may cause thermal decomposition of the sample.

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Source Parameters and Voltages

B

Depending on the ion source installed on the mass spectrometer, different source-dependent parameters are available to optimize.

TurboIonSpray

®

Probe Parameters

The following table shows the recommended operating conditions for the TurboIonSpray probe at three different flow rates. For each flow rate, the Curtain Gas

TM

flow should be as high as possible. The solvent composition used for optimization was 50:50 water:acetonitrile. These conditions represent a starting point from which to optimize the probe. Using an iterative process, optimize the parameters using flow injection analysis to achieve the best signal or signal-to-noise for the compound of interest.

Table B-1 Parameter Optimization for the TurboIonSpray Probe

Parameters

LC flow rate

Nebulizer current

Gas 1 (nebulizer gas)

Gas 2 (heater gas)

Curtain Gas

TM supply

Temperature*

Declustering Potential (DP)

**

5 µL/min to

50 µL/min

Positive: 2 Positive: 2

Negative: –2 Negative: –2

20 psi to 40 psi

Typical Values

200 µL/min

40 psi to 60 psi

1000 µL/min

Positive: 2

Negative: –2

40 psi to 60 psi

0 psi

20 psi

50 psi

30 psi

50 psi

35 psi

0ºC to 200ºC 425ºC to 650ºC

Positive: 70

V

Negative

–70V

Positive: 70 V

Negative –70V

550ºC to 750ºC

Positive: 70 V

Negative –70V

Operational Range

5 µL/min to

3000 µL/min

Positive: 1 to 8

Negative: –1 to 8

0 psi to 90 psi

0 psi to 90 psi

20 psi to 50 psi

Up to 750ºC

Positive: 0 V to 400 V

Negative –400V to 0 V

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Source Parameters and Voltages

Table B-1 Parameter Optimization for the TurboIonSpray Probe (continued)

Parameters

Probe horizontal micrometer setting

5 to 8 5 to 8

Typical Values

5 to 8

Operational Range

0 to 10

* Optimum temperature values depend on the compound and mobile phase composition (higher aqueous content requires higher temperature). Zero (0) means no temperature is applied.

** DP values depends on the compound.

APCI Probe Parameters

Table B-2 Parameter Optimization for the APCI Probe

Parameter

LC flow rate

Gas 2(nebulizer gas)

Curtain Gas

TM

supply

Temperature*

IonSpray

TM

Voltage Floating

Typical Value

1000 µL/min

30

25

400ºC

Declustering Potential (DP)

Positive: 5500

Negative: –4500

Positive: 60 V

Negative: –60 V

Probe vertical micrometer setting

* Temperature value depends on the compound.

Operational Range

200 µL/min to 2000 µL/min

0 to 90

20 to 50

100ºC to 750ºC

Positive: 0 to 5500

Negative: –4500 to 0

Positive: 0 V to 300 V

Negative: –300 V to 0 V

Scale 0 mm to 13 mm

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Source Parameters and Voltages

Parameter Descriptions

Table B-3 Source-Dependent Parameters

Parameter

Ion Source Gas 1

(GS1)

Ion Source Gas 2

(GS2)

Curtain Gas (CUR)

Temperature (TEM)

Description

Controls the nebulizer gas for the TurboIonSpray

®

Operation—Ion Source on page 39

.

probe. Refer to

Principles of

TurboIonSpray probe: Controls the heater gas. The best sensitivity is achieved when the combination of temperature (TEM) and heater gas (GS2) flow rate causes the LC solvent to reach a point at which it is nearly all vaporized. To optimize GS2, increase the flow to obtain the best signal or signal-to-noise ratio. If you see a significant increase in background noise, reduce the value. Too high a gas flow can produce a noisy or unstable signal.

APCI probe: Controls the nebulizer gas.

Refer to

Principles of Operation—Ion Source on page 39

.

Controls the flow of gas to the Curtain Gas

TM

interface. The Curtain Gas interface is located between the curtain plate and the orifice. It prevents ambient air and solvent droplets from entering and contaminating the ion optics, while permitting direction of sample ions into the vacuum chamber by the electrical fields generated between the vacuum interface and the spray needle. Contamination of the ion entrance optics thus reduces Q0 transmission, stability, and sensitivity, and increases background noise.

Maintain the Curtain Gas flow as high as possible without losing sensitivity.

Controls the heat applied to the sample to vaporize it. The optimal temperature is the lowest temperature at which the sample is vaporized completely.

Temperature is applied to both probes simultaneously.

Optimize in increments of 50°C.

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Source Parameters and Voltages

Table B-3 Source-Dependent Parameters (continued)

Parameter

Temperature (TEM) -

TurboIonSpray probe

Temperature (TEM) -

APCI probe

Description

Controls the temperature of the heater gas in the TurboIonSpray probe.

The best sensitivity is achieved when the combination of temperature (TEM) and heater gas (GS2) flow rate causes the LC solvent to reach a point at which it is nearly all vaporized.

As the organic content of the solvent increases, the optimal probe temperature should decrease. With solvents consisting of 100% methanol or acetonitrile, the probe performance may optimize as low as 300°C. Aqueous solvents consisting of 100% water at flows of approximately 1000 µL/min require a maximum probe temperature of 750°C.

If the temperature is set too low, then vaporization is incomplete and large, visible droplets are expelled into the ion source housing.

If the temperature is set too high, solvent may vaporize prematurely at the TurboIonSpray probe tip, especially if the probe is set too low (5 mm to 13 mm).

Controls the temperature of the APCI probe.

As the organic content of the solvent increases, the optimal probe temperature should decrease. With solvents consisting of 100% methanol or acetonitrile the probe performance may optimize at temperatures as low as 400°C at flow rates of 1000 µL/min.

Aqueous solvents consisting of 100% water set at flows of approximately 2000 µL/min require a minimum probe temperature of 700°C.

If the temperature is set too low, then vaporization is incomplete and large, visible droplets are expelled into the ion source housing.

If the temperature is set too high, thermal degradation of the sample occurs.

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Source Parameters and Voltages

Table B-3 Source-Dependent Parameters (continued)

Parameter

IonSpray Voltage

Floating (ISVF)

Interface Heater (ihe)

Description

The IonSpray

TM

voltage is used for both the TurboIonSpray and APCI probes. It is applied to both probes simultaneously.

TurboIonSpray probe: Controls the voltage applied to the sprayer, which ionizes the sample in the ion source. It depends on the polarity, and affects the stability of the spray and the sensitivity.

APCI probe: Controls the current applied to the corona discharge needle. The discharge ionizes solvent molecules, which in turn ionize the sample molecules. The current usually optimizes over a broad range. To optimize in positive mode, start at a value of 5500 V and decrease in increments of 500 V to achieve the best signal or signal-to-noise ratio.

In negative mode, start at a value of –4500 V, and increase in increments of 500 V.

This parameter is always on for TripleTOF

®

systems.

The ihe parameter turns the interface heater on and off. Heating the interface helps maximize the ion signal and prevents contamination of the ion optics. Unless the compound the user is analyzing is extremely fragile, we recommend that the user heats the interface.

Probe Position

The position of the probe can affect the sensitivity of the analysis. Refer to

Ion Source Optimization on page 17

for more information on how to optimize the position of the probe.

Solvent Composition

The standard concentration of ammonium formate or ammonium acetate is from 2 mmol/L to 10 mmol/L for positive ions and 2 mmol/L to 50 mmol/L for negative ions. The concentration of organic acids is 0.1% to 0.5% by volume for the TurboIonSpray

®

probe and 0.1% to 2.0% by volume for the APCI probe.

Commonly used solvents are:

• Acetonitrile

• Methanol

• Propanol

• Water

Commonly used modifiers are:

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Source Parameters and Voltages

• Acetic acid

• Formic acid

• Ammonium formate

• Ammonium acetate

The following modifiers are not commonly used because they complicate the spectrum with their ion mixtures and cluster combinations. They might also suppress the strength of the target compound ion signal:

• Triethyl amine (TEA)

• Sodium phosphate

• Trifluoroacetic acid (TFA)

• Sodium dodecyl sulfate

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Consumables and Spares

C

The following tables list parts included in the consumable parts kit (PN 1005603) as well as spare parts for the

DuoSpray

TM ion source.

Table C-1 Orderable Parts

Part Number

016316

016325

025388

025392

027471

1005601

1005602

025348

026626

Description

PEEK tubing, Red, 1/16 inch o.d. × 0.005 bore

PEEK fitting, Brown, 10-32 × 1/16 inch

Electrode, Nebulizer

Electrode, TurboIonSpray

PEEK Graph-tite fitting, Black, 1/16 inch

PEEK tubing kit to connect to TurboIonSpray

® probe, 30 cm

1

PEEK tubing kit to connect to APCI probe, 45 cm 1

1

2

PEEK union in probe

Spring for probe

1

1

5

1

Quantity

100 cm

Table C-2 Spares

Part Number

1006177

1006174

027497

027013

Description

APCI Corona discharge needle tip

APCI Corona discharge needle rod

Gold-plated spring for HV connection

Spring for corona discharge needle

Quantity

1

1

1

1

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Revision History

Document Number

D5007928 A

D5007928 B

D5007928 C

D5007928 D

RUO-IDV-05-0783-A

Description of Change

First release of document.

Removed CDS information.

Updated the testing procedure.

Updated for the TripleTOF

®

4600 system.

Removed the turbo heater replacement procedure.

Applied new template. Updated for the TripleTOF

®

6600 system.

Date

June 2010

January 2011

March 2011

June 2012

June 2014

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Index

A

adjusting, electrode tip 31

APCI probe

components 9

gas parameters and Curtain Gas flow, optimizing 23

ionization region 44

optimizing probe temperature 25

overview 8

parameters 47

parts of 30

positioning 24

principles of operation 40

probe position, optimizing 24

set up the system 19, 22

atmospheric pressure chemical ionization, described 42

C

cleaning

electrode tube 29

probes 27

components

APCI probe 9

TurboIonSpray probe 8

connections

gas and electrical, ion source 9

corona discharge needle tip, replacing 32

corona discharge, causes of 21

Curtain Gas parameter

defined 48

E

electrode tip

adjusting 31

parts of 32

electrode tube

cleaning 29

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G

cleaning frequency 29

I

gas parameters

optimizing 21, 23

GS1 parameter

optimizing 21

GS2 parameter

defined 48

optimizing 21

ihe parameter, defined 50

installing

ion source 14

ion evaporation, described 40

ion source

connections 9

identification of in the software 12 installation, required materials 12

installing 14

ion source sense circuit, described 10

optimizing 17

preventative maintenance 26

removing 28

required materials, maintenance 26

running methods 19, 22 set up the system 19, 22

source exhaust system, described 11

ISVF parameter, defined 50

M

maintenance

ion source, preventative 26

methods

running 19, 22

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N

nebulizer gas

optimizing 21

O

optimize

TurboIonSpray probe position 20

optimizing

APCI probe position 24

APCI probe temperature 25

gas parameters and Curtain Gas flow 23

GS1 parameter 21

GS2 parameter 21

ion source 17

nebulizer gas 21 turbo heater temperature 21

TurboIonSpray probe 18

voltages 21

organic content and probe temperature 19

P

parameters

APCI probe 47

parameters, optimizing 21

set the starting conditions 19, 23

parts components

probes

APCI probe temperature, optimizing 25

APCI probe, overview 8

cleaning 27

flow rate and temperature 19

optimizing the TurboIonSpray probe 18

optimizing voltages 21

parameters 46

parts of APCI probe 30

removing 29

selecting 7

turbo heater temperature, optimizing 21

TurboIonSpray probe, overview 8

use of 5

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Index

R

removing

ion source 28

probes from the ion source 29

replacing

corona discharge needle tip 32

sample tubing 35

required materials

ion source installation 12

S

sample stream injection 17

sample tubing

blockages 35 replacing 35

sample tubing, cleaning 28

sense circuit, ion source 10

software

identifying the ion source 12

solvents

composition of 50

source exhaust system, described 11

T

technical support requirements

TEM parameter, defined 48

temperature

APCI probe temperature, optimizing 25

turbo heater temperature, optimizing 21

TIS TurboIonSpray probe

TurboIonSpray probe

components 8

optimize the probe position 20

optimizing 18

overview 8

parameters 46

principles of operation 39

turbo heater temperature, optimizing 21 voltages, optimizing 21

V

voltages, optimizing 21

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